Creation of Human Skin Equivalents for the In vitro Study of Angiogenesis in Wound Healing

In our efforts aimed at studying the cellular responses to injury, including the angiogenesis of wound healing, we have developed a novel three-dimensional (3D) skin equivalent that is comprised of multiple cell types found in normal human skin or chronic wound beds. The in vitro model contains a microvascular component within the dermis-like extracellular matrix and possesses an intact epithelial covering comprised of skin-derived epithelial cells. Capillary endothelial cells can be labeled with fluorescent vital tracers prior to being embedded within a 3D matrix and overlaid with a monolayer of keratinocytes (normal or transformed). Once embedded in the matrix, the endothelial cells demonstrate capillary-like tube formation mimicking the microvasculature of true skin. Angiogenesis and the reepithelialization, which occur in response to injury and during wound healing, can be quantified using fluorescence-based and bright-field digital imaging microscopic, biochemical, or molecular approaches.